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Phospholipid oxidation generates potent anti-inflammatory lipid mediators that mimic structurally related pro-resolving eicosanoids by activating Nrf2.

Bretscher P, Egger J, Shamshiev A, Trötzmüller M, Köfeler H, Carreira EM, Kopf M, Freigang S - EMBO Mol Med (2015)

Bottom Line: While the ability of OxPL to modulate biological processes is increasingly recognized, the nature of the biologically active OxPL species and the molecular mechanisms underlying their signaling remain largely unknown.Our study defines epoxycyclopentenones as potent anti-inflammatory lipid mediators that mimic the signaling of endogenous, pro-resolving prostanoids by activating the transcription factor nuclear factor E2-related factor 2 (Nrf2).Using a library of OxPL variants, we identified a synthetic OxPL derivative, which alleviated endotoxin-induced lung injury and inhibited development of pro-inflammatory T helper (Th) 1 cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Health Sciences, ETH Zurich, Zurich, Switzerland.

No MeSH data available.


Related in: MedlinePlus

Cyclo-EC is an EC variant with superior anti-inflammatory bioactivity in vitro and in vivoChemical structure of the cyclic EC analog cyclo-EC.Quantification of IL-6 and IL-12 secretion by BMDC treated with 250 nM cEC or EC for 60 min before stimulation with R837 (5 μg/ml; 18 h). Bars represent mean ± SEM from one of three independent experiments. **P < 0.01; ***P < 0.001; one-way ANOVA adjusted by Dunnett's multiple comparisons test.mRNA expression of the Nrf2 targets Hmox1 and Nqo1 normalized to G6pdx expression. BMDCs were treated for 60 min with 500 nM cEC or DPPC followed by R837 stimulation (5 μg/ml) for 3 h. Data (mean ± SD) are representative of three independent experiments. *P < 0.05; **P < 0.01; unpaired two-tailed t-test.ΔEC50 values of the indicated lipids and OxPL shown relative to that of EC as determined in (E).Dose–response curves showing the modulation of R837-induced (5 μg/ml; 18 h) IL-12 secretion in BMDCs by prior treatment with the indicated OxPL derivatives for 1 h.mRNA quantification of the indicated chemokines relative to G6pdx expression. BMDC were pretreated for 60 min with 500 nM cEC or the variant BisRed prior to R837 stimulation (5 μg/ml) for 3 h. Data (mean ± SD) are representative of three independent experiments. *P < 0.05; **P < 0.01; unpaired two-tailed t-test.Quantification and characterization of cellular infiltrates in BAL. Groups of six C57BL/6 mice were pretreated i.t. with 50 μg cEC, EC, or BisRed 24 and 2 h before challenge with 150 ng/g LPS in the presence of 800 μg/g D-galactosamine. BAL was harvested 4 h after LPS injection, and inflammatory cells were characterized by FACS analysis. **P < 0.01; ****P < 0.0001; one-way ANOVA with Sidak's multiple comparisons test.Comparison of the capacity of cEC, EC, and PECPC (1 μM) to license splenic dendritic cells to polarize naïve CD4 T cells into IFN-γ-producing (Th1) and IL-4-producing (Th2) effector cells. Data (mean ± SD) are representative of two independent experiments. One-way ANOVA adjusted by Dunnett's multiple comparisons test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.
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fig07: Cyclo-EC is an EC variant with superior anti-inflammatory bioactivity in vitro and in vivoChemical structure of the cyclic EC analog cyclo-EC.Quantification of IL-6 and IL-12 secretion by BMDC treated with 250 nM cEC or EC for 60 min before stimulation with R837 (5 μg/ml; 18 h). Bars represent mean ± SEM from one of three independent experiments. **P < 0.01; ***P < 0.001; one-way ANOVA adjusted by Dunnett's multiple comparisons test.mRNA expression of the Nrf2 targets Hmox1 and Nqo1 normalized to G6pdx expression. BMDCs were treated for 60 min with 500 nM cEC or DPPC followed by R837 stimulation (5 μg/ml) for 3 h. Data (mean ± SD) are representative of three independent experiments. *P < 0.05; **P < 0.01; unpaired two-tailed t-test.ΔEC50 values of the indicated lipids and OxPL shown relative to that of EC as determined in (E).Dose–response curves showing the modulation of R837-induced (5 μg/ml; 18 h) IL-12 secretion in BMDCs by prior treatment with the indicated OxPL derivatives for 1 h.mRNA quantification of the indicated chemokines relative to G6pdx expression. BMDC were pretreated for 60 min with 500 nM cEC or the variant BisRed prior to R837 stimulation (5 μg/ml) for 3 h. Data (mean ± SD) are representative of three independent experiments. *P < 0.05; **P < 0.01; unpaired two-tailed t-test.Quantification and characterization of cellular infiltrates in BAL. Groups of six C57BL/6 mice were pretreated i.t. with 50 μg cEC, EC, or BisRed 24 and 2 h before challenge with 150 ng/g LPS in the presence of 800 μg/g D-galactosamine. BAL was harvested 4 h after LPS injection, and inflammatory cells were characterized by FACS analysis. **P < 0.01; ****P < 0.0001; one-way ANOVA with Sidak's multiple comparisons test.Comparison of the capacity of cEC, EC, and PECPC (1 μM) to license splenic dendritic cells to polarize naïve CD4 T cells into IFN-γ-producing (Th1) and IL-4-producing (Th2) effector cells. Data (mean ± SD) are representative of two independent experiments. One-way ANOVA adjusted by Dunnett's multiple comparisons test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Mentions: Considering the structural and functional similarity of EC and 15d-PGJ2, we also tested a series of chimeric molecules that combined features of EC and 15d-PGJ2, the synthesis of which is described in detail elsewhere (Egger et al, 2014). In this process, we developed the EC variant cyclo-EC (cEC), which hypothetically could be generated by an intra-molecular nucleophilic attack of the carboxylate-anion on one of the epoxide carbons, thus forming a 6-membered lactone ring (Fig7A) (Egger et al, 2014). Compared to EC, cEC exhibited superior anti-inflammatory capacity, as assessed by inhibition of IL-6 and IL-12 production (Fig7B), but also by the transcriptional regulation of IL-12, IL-6, IL-23, and TNFα (Supplementary Fig S7). The strong induction of several Nrf2 target genes including Hmox1 and Nqo1 indicated that like EC, also cEC triggered Nrf2 signaling to mediate these effects (Fig7C and Supplementary Fig S7). Moreover, cEC showed the most potent inhibition of IL-12 secretion among all fatty acid cyclopentenone OxPL tested (Fig7D and E). Expectedly, cEC also reduced the expression of pro-inflammatory chemokines (Fig7F) and decreased the infiltration of neutrophils into the lungs of LPS-challenged animals (Fig7G). The improved efficacy of cEC was further demonstrated by its superior capacity to modulate the DC-licensed T-cell differentiation in vitro (Fig7H). In particular, we observed that cEC still strongly biased the polarization of naïve T cells at concentrations at which EC only exhibited weak residual activity and no such effects could be detected for 15d-PGJ2 and PECPC. These data not only established cEC as a promising epoxycyclopentenone-derived compound to be further evaluated in the treatment of inflammatory disorders, but also recommended the class of epoxycyclopentenone-containing OxPL as potential basis for future anti-inflammatory therapeutics.


Phospholipid oxidation generates potent anti-inflammatory lipid mediators that mimic structurally related pro-resolving eicosanoids by activating Nrf2.

Bretscher P, Egger J, Shamshiev A, Trötzmüller M, Köfeler H, Carreira EM, Kopf M, Freigang S - EMBO Mol Med (2015)

Cyclo-EC is an EC variant with superior anti-inflammatory bioactivity in vitro and in vivoChemical structure of the cyclic EC analog cyclo-EC.Quantification of IL-6 and IL-12 secretion by BMDC treated with 250 nM cEC or EC for 60 min before stimulation with R837 (5 μg/ml; 18 h). Bars represent mean ± SEM from one of three independent experiments. **P < 0.01; ***P < 0.001; one-way ANOVA adjusted by Dunnett's multiple comparisons test.mRNA expression of the Nrf2 targets Hmox1 and Nqo1 normalized to G6pdx expression. BMDCs were treated for 60 min with 500 nM cEC or DPPC followed by R837 stimulation (5 μg/ml) for 3 h. Data (mean ± SD) are representative of three independent experiments. *P < 0.05; **P < 0.01; unpaired two-tailed t-test.ΔEC50 values of the indicated lipids and OxPL shown relative to that of EC as determined in (E).Dose–response curves showing the modulation of R837-induced (5 μg/ml; 18 h) IL-12 secretion in BMDCs by prior treatment with the indicated OxPL derivatives for 1 h.mRNA quantification of the indicated chemokines relative to G6pdx expression. BMDC were pretreated for 60 min with 500 nM cEC or the variant BisRed prior to R837 stimulation (5 μg/ml) for 3 h. Data (mean ± SD) are representative of three independent experiments. *P < 0.05; **P < 0.01; unpaired two-tailed t-test.Quantification and characterization of cellular infiltrates in BAL. Groups of six C57BL/6 mice were pretreated i.t. with 50 μg cEC, EC, or BisRed 24 and 2 h before challenge with 150 ng/g LPS in the presence of 800 μg/g D-galactosamine. BAL was harvested 4 h after LPS injection, and inflammatory cells were characterized by FACS analysis. **P < 0.01; ****P < 0.0001; one-way ANOVA with Sidak's multiple comparisons test.Comparison of the capacity of cEC, EC, and PECPC (1 μM) to license splenic dendritic cells to polarize naïve CD4 T cells into IFN-γ-producing (Th1) and IL-4-producing (Th2) effector cells. Data (mean ± SD) are representative of two independent experiments. One-way ANOVA adjusted by Dunnett's multiple comparisons test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.
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fig07: Cyclo-EC is an EC variant with superior anti-inflammatory bioactivity in vitro and in vivoChemical structure of the cyclic EC analog cyclo-EC.Quantification of IL-6 and IL-12 secretion by BMDC treated with 250 nM cEC or EC for 60 min before stimulation with R837 (5 μg/ml; 18 h). Bars represent mean ± SEM from one of three independent experiments. **P < 0.01; ***P < 0.001; one-way ANOVA adjusted by Dunnett's multiple comparisons test.mRNA expression of the Nrf2 targets Hmox1 and Nqo1 normalized to G6pdx expression. BMDCs were treated for 60 min with 500 nM cEC or DPPC followed by R837 stimulation (5 μg/ml) for 3 h. Data (mean ± SD) are representative of three independent experiments. *P < 0.05; **P < 0.01; unpaired two-tailed t-test.ΔEC50 values of the indicated lipids and OxPL shown relative to that of EC as determined in (E).Dose–response curves showing the modulation of R837-induced (5 μg/ml; 18 h) IL-12 secretion in BMDCs by prior treatment with the indicated OxPL derivatives for 1 h.mRNA quantification of the indicated chemokines relative to G6pdx expression. BMDC were pretreated for 60 min with 500 nM cEC or the variant BisRed prior to R837 stimulation (5 μg/ml) for 3 h. Data (mean ± SD) are representative of three independent experiments. *P < 0.05; **P < 0.01; unpaired two-tailed t-test.Quantification and characterization of cellular infiltrates in BAL. Groups of six C57BL/6 mice were pretreated i.t. with 50 μg cEC, EC, or BisRed 24 and 2 h before challenge with 150 ng/g LPS in the presence of 800 μg/g D-galactosamine. BAL was harvested 4 h after LPS injection, and inflammatory cells were characterized by FACS analysis. **P < 0.01; ****P < 0.0001; one-way ANOVA with Sidak's multiple comparisons test.Comparison of the capacity of cEC, EC, and PECPC (1 μM) to license splenic dendritic cells to polarize naïve CD4 T cells into IFN-γ-producing (Th1) and IL-4-producing (Th2) effector cells. Data (mean ± SD) are representative of two independent experiments. One-way ANOVA adjusted by Dunnett's multiple comparisons test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.
Mentions: Considering the structural and functional similarity of EC and 15d-PGJ2, we also tested a series of chimeric molecules that combined features of EC and 15d-PGJ2, the synthesis of which is described in detail elsewhere (Egger et al, 2014). In this process, we developed the EC variant cyclo-EC (cEC), which hypothetically could be generated by an intra-molecular nucleophilic attack of the carboxylate-anion on one of the epoxide carbons, thus forming a 6-membered lactone ring (Fig7A) (Egger et al, 2014). Compared to EC, cEC exhibited superior anti-inflammatory capacity, as assessed by inhibition of IL-6 and IL-12 production (Fig7B), but also by the transcriptional regulation of IL-12, IL-6, IL-23, and TNFα (Supplementary Fig S7). The strong induction of several Nrf2 target genes including Hmox1 and Nqo1 indicated that like EC, also cEC triggered Nrf2 signaling to mediate these effects (Fig7C and Supplementary Fig S7). Moreover, cEC showed the most potent inhibition of IL-12 secretion among all fatty acid cyclopentenone OxPL tested (Fig7D and E). Expectedly, cEC also reduced the expression of pro-inflammatory chemokines (Fig7F) and decreased the infiltration of neutrophils into the lungs of LPS-challenged animals (Fig7G). The improved efficacy of cEC was further demonstrated by its superior capacity to modulate the DC-licensed T-cell differentiation in vitro (Fig7H). In particular, we observed that cEC still strongly biased the polarization of naïve T cells at concentrations at which EC only exhibited weak residual activity and no such effects could be detected for 15d-PGJ2 and PECPC. These data not only established cEC as a promising epoxycyclopentenone-derived compound to be further evaluated in the treatment of inflammatory disorders, but also recommended the class of epoxycyclopentenone-containing OxPL as potential basis for future anti-inflammatory therapeutics.

Bottom Line: While the ability of OxPL to modulate biological processes is increasingly recognized, the nature of the biologically active OxPL species and the molecular mechanisms underlying their signaling remain largely unknown.Our study defines epoxycyclopentenones as potent anti-inflammatory lipid mediators that mimic the signaling of endogenous, pro-resolving prostanoids by activating the transcription factor nuclear factor E2-related factor 2 (Nrf2).Using a library of OxPL variants, we identified a synthetic OxPL derivative, which alleviated endotoxin-induced lung injury and inhibited development of pro-inflammatory T helper (Th) 1 cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Health Sciences, ETH Zurich, Zurich, Switzerland.

No MeSH data available.


Related in: MedlinePlus